# this cell is tagged as parameters for `papermill` parameterization
altair_config = None
nipah_config = None

codon_variants_file = None

CHO_corr_plot_save = None
CHO_EFNB2_indiv_plot_save = None
CHO_EFNB3_indiv_plot_save = None

histogram_plot = None
func_scores_plot = None
uniq_barcodes_per_lib_df = None

# Parameters
nipah_config = "nipah_config.yaml"
altair_config = "data/custom_analyses_data/theme.py"
codon_variants_file = "results/variants/codon_variants.csv"
CHO_corr_plot_save = "results/images/CHO_corr_plot_save.html"
CHO_EFNB2_indiv_plot_save = "results/images/CHO_EFNB2_all_corrs.html"
CHO_EFNB3_indiv_plot_save = "results/images/CHO_EFNB3_all_corrs.html"
histogram_plot = "results/images/variants_histogram.html"
func_scores_plot = "results/images/func_scores_distribution.html"
uniq_barcodes_per_lib_df = "results/tables/uniq_barcodes_per_lib_df.csv"

import math
import os
import re
import altair as alt

import numpy as np

import pandas as pd

import scipy.stats

import Bio.SeqIO
import yaml
from Bio import AlignIO
from Bio import PDB
from Bio.Align import PairwiseAligner
from collections import Counter

# allow more rows for Altair
_ = alt.data_transformers.disable_max_rows()

if os.getcwd() == '/fh/fast/bloom_j/computational_notebooks/blarsen/2023/Nipah_Malaysia_RBP_DMS/':
    pass
    print("Already in correct directory")
else:
    os.chdir("/fh/fast/bloom_j/computational_notebooks/blarsen/2023/Nipah_Malaysia_RBP_DMS/")
    print("Setup in correct directory")

Setup in correct directory

if histogram_plot is None:
    altair_config = 'data/custom_analyses_data/theme.py'
    nipah_config = 'nipah_config.yaml'
    codon_variants_file = 'results/variants/codon_variants.csv'
#CHO_corr_plot_save
#CHO_EFNB3_corr_plot_save
#CHO_EFNB2_indiv_plot_save
#CHO_EFNB3_indiv_plot_save

if altair_config:
    with open(altair_config, 'r') as file:
        exec(file.read())

with open(nipah_config) as f:
    config = yaml.safe_load(f)

with open('data/func_effects_config.yml') as y:
    config_func = yaml.safe_load(y)

cho_efnb2_low_selections = config_func['avg_func_effects']['CHO_EFNB2_low']['selections']
LibA_CHO_EFNB2 = [selection + '_func_effects.csv' for selection in cho_efnb2_low_selections if 'LibA' in selection and 'LibB' not in selection]
LibB_CHO_EFNB2 = [selection + '_func_effects.csv' for selection in cho_efnb2_low_selections if 'LibB' in selection and 'LibA' not in selection]

cho_efnb3_low_selections = config_func['avg_func_effects']['CHO_EFNB3_low']['selections']
LibA_CHO_EFNB3 = [selection + '_func_effects.csv' for selection in cho_efnb3_low_selections if 'LibA' in selection and 'LibB' not in selection]
LibB_CHO_EFNB3 = [selection + '_func_effects.csv' for selection in cho_efnb3_low_selections if 'LibB' in selection and 'LibA' not in selection]

path = 'results/func_effects/by_selection/'
def process_func_selections(library,library_name):
    df_list = []
    clock = 1
    for file_name in library:
        file_path = os.path.join(path, file_name)

        fixed_name = file_name.replace('_func_effects.csv', '')
        
        # Read the current CSV file
        func_scores = pd.read_csv(file_path)
        
        func_scores_renamed = func_scores.rename(columns={'functional_effect': f'functional_effect_{clock}','times_seen': f'times_seen_{clock}'})
        func_scores_renamed.drop(['latent_phenotype_effect'],axis=1,inplace=True)
        
        # Append the dataframe to the list
        df_list.append(func_scores_renamed)
        clock = clock + 1
    
    # Merge all dataframes on 'site' and 'mutant'
    merged_df = df_list[0]
    for df in df_list[1:]:
        merged_df = pd.merge(merged_df, df, on=['site', 'mutant','wildtype'], how='outer')
    
    #Calculate median values
    lib_columns_func = [col for col in merged_df.columns if 'functional_effect' in col]
    merged_df[f'median_effect_{library_name}'] = merged_df[lib_columns_func].median(axis=1)
    lib_columns_times_seen = [col for col in merged_df.columns if 'times_seen' in col]
    merged_df[f'median_times_seen_{library_name}'] = merged_df[lib_columns_times_seen].median(axis=1)
    #Now drop columns
    lib_columns = [col for col in merged_df.columns if re.search(r'_\d+', col)]
    merged_df = merged_df.drop(columns=lib_columns)
    return merged_df

A_selections_E2 = process_func_selections(LibA_CHO_EFNB2,'LibA')
B_selections_E2 = process_func_selections(LibB_CHO_EFNB2,'LibB')

A_selections_E3 = process_func_selections(LibA_CHO_EFNB3,'LibA')
B_selections_E3 = process_func_selections(LibB_CHO_EFNB3,'LibB')

def merge_selections(A_selections,B_selections):
    merged_selections = pd.merge(A_selections,B_selections,on=['wildtype','site','mutant'],how='inner')
    
    #make one times seen column for slider
    lib_columns_times_seen = [col for col in merged_selections.columns if 'times_seen' in col]
    merged_selections['times_seen'] = merged_selections[lib_columns_times_seen].median(axis=1)
    return merged_selections

CHO_EFNB2_merged = merge_selections(A_selections_E2,B_selections_E2)
CHO_EFNB2_merged['cell_type'] = 'CHO-EFNB2'
CHO_EFNB3_merged = merge_selections(A_selections_E3,B_selections_E3)
CHO_EFNB3_merged['cell_type'] = 'CHO-EFNB3'

both_selections = pd.concat([CHO_EFNB2_merged, CHO_EFNB3_merged])

def make_chart_median(df,title):
    slider = alt.binding_range(min=1, max=25, step=1, name="times_seen")
    selector = alt.param(name="SelectorName", value=1, bind=slider)
    
    empty = []
    variant_selector = alt.selection_point(
        on="mouseover",
        empty=False,
        nearest=True,
        fields=["site","mutant"],
        value=1
    )
  
    df = df[
        (df['median_effect_LibA'].notna()) &
        (df['median_effect_LibB'].notna())
    ]
    size = df.shape[0]
    

    for selection in ['CHO-EFNB2','CHO-EFNB3']:
        print(selection)
        tmp_df = df[df['cell_type'] == selection] 
        slope, intercept, r_value, p_value, std_err = scipy.stats.linregress(df[f'median_effect_LibA'], df[f'median_effect_LibB'])
        r_value = float(r_value)
        print(f'{r_value:.2f}')
        
        chart = alt.Chart(tmp_df,title=f'Entry in {selection} cells').mark_point().encode(
            x=alt.X('median_effect_LibA',title='LibA Cell Entry'),
            y=alt.Y('median_effect_LibB',title='LibB Cell Entry'),
            tooltip=['site','wildtype','mutant','times_seen'],
            size=alt.condition(variant_selector, alt.value(100),alt.value(15)),
            color=alt.condition(alt.datum.times_seen < selector, alt.value('transparent'), alt.value('black')),
            opacity=alt.condition(variant_selector, alt.value(1),alt.value(0.2)),
        )
        empty.append(chart)
    combined_effect_chart = alt.hconcat(*empty).resolve_scale(y='shared', x='shared', color='independent').add_params(variant_selector,selector)
    return combined_effect_chart

CHO_EFNB2_corr_plot = make_chart_median(both_selections,'CHO-EFNB2')
CHO_EFNB2_corr_plot.display()
if histogram_plot is not None:
    CHO_EFNB2_corr_plot.save(CHO_corr_plot_save)

CHO-EFNB2
0.92
CHO-EFNB3
0.92

def plot_corr_heatmap(df):
    empty_chart = []
    
    for cell in list(df['cell_type'].unique()):
        tmp_df = df[df['cell_type'] == cell]
        chart = alt.Chart(tmp_df,title=f'{cell}').mark_rect().encode(
            x=alt.X('median_effect_LibA',title='Library A').bin(maxbins=50), #axis=alt.Axis(values=[-4,-1,0,1])
            y=alt.Y('median_effect_LibB',title='Library B').bin(maxbins=50), #,axis=alt.Axis(values=[-4,-1,0,1])
            color=alt.Color('count()',title='Count').scale(scheme='greenblue'),
            #tooltip=['effect','binding_median']
        )
        empty_chart.append(chart)
    
    combined_chart = alt.hconcat(*empty_chart,title=alt.Title('Correlation between binding and entry')).resolve_scale(y='shared',x='shared',color='shared')
    return combined_chart

entry_binding_corr_heatmap = plot_corr_heatmap(both_selections)
entry_binding_corr_heatmap.display()
#entry_binding_corr_heatmap.save(entry_binding_corr_heatmap)

#path = 'results/func_effects/by_selection/'
def process_individ_selections(library):
    df_list = []
    clock = 1
    for file_name in library:
        file_path = os.path.join(path, file_name)
        print(f"Processing file: {file_path}")
        
        #fixed_name = file_name.replace('_func_effects.csv', '')
        
        # Read the current CSV file
        func_scores = pd.read_csv(file_path)
        #display(func_scores.head(2))
        #func_scores = func_scores[func_scores['times_seen'] >= config['func_times_seen_cutoff']]
        func_scores_renamed = func_scores.rename(columns={'functional_effect': f'functional_effect_{clock}','times_seen': f'times_seen_{clock}'})
        func_scores_renamed.drop(['latent_phenotype_effect'],axis=1,inplace=True)
        
        # Append the dataframe to the list
        df_list.append(func_scores_renamed)
        clock = clock + 1
    
    # Merge all dataframes on 'site' and 'mutant'
    merged_df = df_list[0]
    for df in df_list[1:]:
        merged_df = pd.merge(merged_df, df, on=['site', 'mutant','wildtype'], how='outer')
    # Make list of how many selections are done for later correlation plots
    lib_size = len(library)
    number_list = [i for i in range(1, lib_size+1)]
    return merged_df,number_list

CHO_EFNB2_indiv,lib_size_EFNB2 = process_individ_selections(LibA_CHO_EFNB2+LibB_CHO_EFNB2)
CHO_EFNB3_indiv,lib_size_EFNB3 = process_individ_selections(LibA_CHO_EFNB3+LibB_CHO_EFNB3)

def make_chart(df,number_list):
    empty_list = []
    for i in number_list:
        other_empty_list = []
        for j in number_list:
            df = df[
                (df[f'times_seen_{i}'] >= config['func_times_seen_cutoff']) & 
                (df[f'times_seen_{j}'] >= config['func_times_seen_cutoff']) &
                (df[f'functional_effect_{i}'].notna()) &
                (df[f'functional_effect_{j}'].notna())
            ]
            #slope, intercept, r_value, p_value, std_err = scipy.stats.linregress(df[f'functional_effect_{i}'], df[f'functional_effect_{j}'])
            #r_value = float(r_value)
            #print(f'{r_value:.2f}')
            chart = alt.Chart(df).mark_circle(size=10, color='black', opacity=0.2).encode(
                x=alt.X(f'functional_effect_{i}'),
                y=alt.Y(f'functional_effect_{j}'),
                tooltip=['site','wildtype','mutant'],
            ).properties(
                height=alt.Step(10),
                width=alt.Step(10)
            )
            other_empty_list.append(chart)
        combined_effect_chart = alt.hconcat(*other_empty_list).resolve_scale(y='shared', x='shared', color='independent')
        empty_list.append(combined_effect_chart)
    final_combined_chart = alt.vconcat(*empty_list).resolve_scale(y='shared', x='shared', color='independent')
    return final_combined_chart

CHO_EFNB2_indiv_plot = make_chart(CHO_EFNB2_indiv,lib_size_EFNB2)
#CHO_EFNB2_indiv_plot.display()
if histogram_plot is not None:
    CHO_EFNB2_indiv_plot.save(CHO_EFNB2_indiv_plot_save)
CHO_EFNB3_indiv_plot = make_chart(CHO_EFNB3_indiv,lib_size_EFNB3)
if histogram_plot is not None:
    CHO_EFNB3_indiv_plot.save(CHO_EFNB3_indiv_plot_save)

Processing file: results/func_effects/by_selection/LibA-231112-CHO-EFNB2-BA6-nac_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231207-CHO-EFNB2-BA6-1_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231207-CHO-EFNB2-BA6-2_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231207-CHO-EFNB2-BA6-3_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231207-CHO-EFNB2-BA6-pool_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231222-CHO-EFNB2-BA6-nac_diffVSV_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-231112-CHO-EFNB2-BA6-nac_diff_VSV_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-231116-CHO-BA6_PREV_POOL_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-230725-CHO-EFNB3-C6-nac-diffVSV_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-230916-CHO-EFNB2-BA6-nac_diffVSV_func_effects.csv
Processing file: results/func_effects/by_selection/LibA-231024-CHO-EFNB3-C6-nac_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-230720-CHO-C6-nac-VSV_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-230815-CHO-C6-nac_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-230818-CHO-C6-nac_func_effects.csv
Processing file: results/func_effects/by_selection/LibB-231116-CHO-C6_PREV_POOL_func_effects.csv

codon_variants = pd.read_csv(codon_variants_file)
display(codon_variants.head(3))
unique_barcodes_per_library = codon_variants.groupby('library')['barcode'].nunique()

uniq_barcodes_per_lib = pd.DataFrame(unique_barcodes_per_library)
display(uniq_barcodes_per_lib)

# Initialize an empty dictionary to keep track of observed mutations
aa_counts = {}
wildtypes = {}  # Dictionary to keep track of wildtype amino acids for each site

# Function to process each cell, update counts, and record wildtype amino acids
def process_cell(cell):
    if pd.notna(cell):  # Check if cell is not NaN
        substitutions = cell.split()
        for substitution in substitutions:
            if substitution[-1] not in ('*', '-') and substitution[0] not in ('*'):  # Skip if substitution ends with '*' or '-'
                site = substitution[1:-1]
                mutation = substitution[-1]
                wildtype = substitution[0]
                site_mutation = site + mutation
                if site not in wildtypes:
                    wildtypes[site] = wildtype
                if site_mutation in aa_counts:
                    aa_counts[site_mutation] += 1
                else:
                    aa_counts[site_mutation] = 1
empty_mutants = []
empty_percent = []
for lib in ['LibA','LibB']:
    # Apply the function to each cell in the 'aa_substitutions' column
    tmp_df = codon_variants[codon_variants['library'] == lib]
    tmp_df['aa_substitutions'].apply(process_cell)
    
    # Generate all possible combinations excluding the wildtype for each site
    expected_sites = range(1, 533)
    possible_mutations = ['A', 'C', 'D', 'E', 'F', 'G', 'H', 'I', 'K', 'L', 'M', 'N', 'P', 'Q', 'R', 'S', 'T', 'V', 'W', 'Y']
    
    # Adjust expected combinations to exclude the wildtype for each site
    expected_combinations = set()
    for site in expected_sites:
        site_str = str(site)
        if site_str in wildtypes:
            wildtype = wildtypes[site_str]
            for mutation in possible_mutations:
                if mutation != wildtype:  # Exclude the wildtype amino acid
                    expected_combinations.add(site_str + mutation)
    
    # Extract the actual combinations from the counts
    actual_combinations = set(aa_counts.keys())
    
    # Find missing combinations
    missing_combinations = expected_combinations - actual_combinations
    
    # Display results
    print(f"Number of unique site-mutation combinations observed: {len(aa_counts)}")
    print(f"Number of missing combinations (excluding wildtypes): {len(missing_combinations)}")
    print(f"Total possible combinations excluding wildtypes: {len(expected_combinations)}")
    empty_percent.append(len(actual_combinations) / len(expected_combinations))


#empty_percent = empty_percent.reset_index()


uniq_barcodes_per_lib['percent'] = empty_percent
uniq_barcodes_per_lib['percent'] = uniq_barcodes_per_lib['percent'] * 100
uniq_barcodes_per_lib = uniq_barcodes_per_lib.round(2)
uniq_barcodes_per_lib = uniq_barcodes_per_lib.reset_index()
uniq_barcodes_per_lib.to_csv(uniq_barcodes_per_lib_df,index=False)

Number of unique site-mutation combinations observed: 10055
Number of missing combinations (excluding wildtypes): 54
Total possible combinations excluding wildtypes: 10108
Number of unique site-mutation combinations observed: 10080
Number of missing combinations (excluding wildtypes): 29
Total possible combinations excluding wildtypes: 10108

def calculate_fraction(library):
    total_A = codon_variants[codon_variants['library'] == library].shape[0]
    for number in range(5):
        fraction = codon_variants[(codon_variants['library'] == library) & (codon_variants['n_aa_substitutions'] == number)].shape[0]
        print(f'For {library}, the fraction of sites with {number} mutations relative to WT are: {fraction/total_A:.2f}')

calculate_fraction('LibA')
calculate_fraction('LibB')

For LibA, the fraction of sites with 0 mutations relative to WT are: 0.11
For LibA, the fraction of sites with 1 mutations relative to WT are: 0.64
For LibA, the fraction of sites with 2 mutations relative to WT are: 0.22
For LibA, the fraction of sites with 3 mutations relative to WT are: 0.03
For LibA, the fraction of sites with 4 mutations relative to WT are: 0.00
For LibB, the fraction of sites with 0 mutations relative to WT are: 0.11
For LibB, the fraction of sites with 1 mutations relative to WT are: 0.65
For LibB, the fraction of sites with 2 mutations relative to WT are: 0.21
For LibB, the fraction of sites with 3 mutations relative to WT are: 0.03
For LibB, the fraction of sites with 4 mutations relative to WT are: 0.00

def plot_histogram(df):
    df = df.drop(columns=['barcode','target','variant_call_support','codon_substitutions','aa_substitutions','n_codon_substitutions'])
    chart = alt.Chart(df).mark_bar(color='black').encode(
        alt.X("n_aa_substitutions:N",title='# of AA Substitutions'), 
        alt.Y('count()', title='Count',axis=alt.Axis(grid=True)), # count() is a built-in aggregation to count rows in each bin
        column=alt.Column('library',header=alt.Header(title=None, labelFontSize=18))
    )
    return chart

histogram = plot_histogram(codon_variants)
histogram.display()
if histogram_plot is not None:
    histogram.save(histogram_plot)

def pull_in_func_scores(df):
    empty_list = []
    for i in df:
        j = i + '_func_scores.csv'
        tmp_df = pd.read_csv(f'results/func_scores/{j}')
        tmp_df['selection'] = i
        empty_list.append(tmp_df)
        tmp_df = pd.concat(empty_list)
        return tmp_df

e2_func_scores_df = pull_in_func_scores(cho_efnb2_low_selections)
e2_func_scores_df['cell_type'] = 'CHO-EFNB2'
e3_func_scores_df = pull_in_func_scores(cho_efnb3_low_selections)
e3_func_scores_df['cell_type'] = 'CHO-EFNB3'

#Make combined dataframe of cell entry data
merged_func_scores = pd.concat([e2_func_scores_df,e3_func_scores_df])

def classify_mutation(row):
    if isinstance(row['aa_substitutions'], str) and '*' in row['aa_substitutions']:
        return 'stop'
    elif row['n_aa_substitutions'] == 0:
        if row['n_codon_substitutions'] >= 1:
            return 'synonymous'
        else:
            return 'wildtype'
    elif row['n_aa_substitutions'] == 1:
        return '1 nonsynonymous'
    elif row['n_aa_substitutions'] >= 2:
        return '>2 nonsynonymous'

# Apply the function to each row in the dataframe to create the new column
merged_func_scores['mutation_class'] = merged_func_scores.apply(classify_mutation, axis=1)

result_df = merged_func_scores.groupby(['barcode','cell_type']).agg(
    func_score=('func_score', 'median'),
    mutation_class=('mutation_class','first')
).reset_index()

tmp = result_df.groupby(['mutation_class','cell_type'])['func_score'].median().reset_index()
tmp = tmp.rename(columns={'func_score':'median_func_score'})

result_df = result_df.merge(tmp,on=['mutation_class','cell_type'],how='left')
display(result_df)

def plot_func_score_distribution(df):
    custom_sort = ['wildtype', 'synonymous', '1 nonsynonymous', '>2 nonsynonymous', 'stop']
    empty_charts = []
    for cell_idx,target_cell in enumerate(['CHO-EFNB2','CHO-EFNB3']):
        charts = []
        first_df = df[df['cell_type'] == target_cell]
        for idx, subset in enumerate(custom_sort):
            tmp_df = first_df[first_df['mutation_class'] == subset]
            
            is_last_plot = idx == len(custom_sort) - 1
            x_axis = alt.Axis(labelAngle=-90, titleFontSize=10,tickCount=3, values=[-10, -5, 0],
                              title="Functional Score" if is_last_plot else None, 
                              labels=True if is_last_plot else False)  # Only show labels for the last plot
    
            first_plot_column = cell_idx == 0
            y_axis = alt.Axis(labelAngle=0,titleAngle=0,title=subset if first_plot_column else None,domain=False,ticks=False,labels=False,titleX=-10,titleAlign='right')
            
            
            chart = alt.Chart(tmp_df,title=(target_cell if idx == 0 else "")).mark_area(color='black').encode(
                x=alt.X('func_score', bin=alt.Bin(step=0.4), axis=x_axis),
                y=alt.Y('count()', title=subset, axis=y_axis),#alt.Axis(domain=False, ticks=False, labels=False)),
                color=alt.Color('median_func_score',title='Median Functional Score',scale=alt.Scale(scheme='greenblue')),
                #row=alt.Row('mutation_class', title=None, sort=custom_sort, header=alt.Header(title=None)),
                #column=alt.Column('cell_type'),
            ).properties(width=100, height=50)
    
            charts.append(chart)
        combined_muts_chart = alt.vconcat(*charts,spacing=0).resolve_scale(y='independent',x='shared',color='shared')
        empty_charts.append(combined_muts_chart)
    # Combine charts using vertical concatenation, adjusting scales and configuration as needed
    combined_chart = alt.hconcat(*empty_charts, spacing=0).resolve_scale(
        y='independent', x='shared', color='shared'
    ).configure_view(
        stroke=None
    ).configure_axis(
        grid=False
    ).configure_title(
        anchor='middle',  # Anchors the title to the start of the chart
        offset=5,  # Adjusts the distance between the title and the chart
        fontSize=16,  # Adjusts the font size of the title
        #dx=5,  # Shifts the title horizontally (use negative value to shift left)
        #dy=-5  # Shifts the title vertically (use negative value to shift up)
    )
    
    return combined_chart

tmp_img = plot_func_score_distribution(result_df)
tmp_img.display()    
if histogram_plot is not None:
    tmp_img.save(func_scores_plot)

	target	library	barcode	variant_call_support	codon_substitutions	aa_substitutions	n_codon_substitutions	n_aa_substitutions
0	gene	LibA	AAAAAAAAAAAAAGAA	5	ACC461ACT ATC475AGC	I475S	2	1
1	gene	LibA	AAAAAAAAAAACCCAT	36	GCG16GAG CAG23GAG	A16E Q23E	2	2
2	gene	LibA	AAAAAAAAAAAGTTTC	6	TAC319CCC	Y319P	1	1

	barcode	cell_type	func_score	mutation_class	median_func_score
0	AAAAAAAAAAGACCCG	CHO-EFNB2	-2.20700	>2 nonsynonymous	-1.8130
1	AAAAAAAAAAGACCCG	CHO-EFNB3	0.03215	>2 nonsynonymous	-3.0560
2	AAAAAAAAACCTATAG	CHO-EFNB2	-0.17420	1 nonsynonymous	-0.6460
3	AAAAAAAAACCTATAG	CHO-EFNB3	-0.85940	1 nonsynonymous	-0.9497
4	AAAAAAAAATCCTACG	CHO-EFNB2	-0.86490	1 nonsynonymous	-0.6460
...	...	...	...	...	...
128340	TTTTTTCGATGAACGA	CHO-EFNB3	-4.15400	>2 nonsynonymous	-3.0560
128341	TTTTTTGCCAAGTGAA	CHO-EFNB2	-0.50200	>2 nonsynonymous	-1.8130
128342	TTTTTTTAAGACTACA	CHO-EFNB3	-2.23100	1 nonsynonymous	-0.9497
128343	TTTTTTTACTCGAATG	CHO-EFNB2	-0.46220	1 nonsynonymous	-0.6460
128344	TTTTTTTACTCGAATG	CHO-EFNB3	1.59500	1 nonsynonymous	-0.9497

This script calculates the correlations between different cell entry selections. Either the median of all LibA selections vs the median of all LibB selections, OR all selections for a specific condition.¶

Calculate correlations for LibA and LibB for CHO-EFNB2 cell entry selections¶

Make correlations between individual selections¶

Now make histogram of variants¶

Find which sites are present, and which are missing¶

Find distribution of functional scores¶